A large and growing corpus of scientific literature is devoted to the key role that mitochondria play in aging. An even larger research effort is devoted to the study of oxidative stress (particularly of mitochondrial origin) and its role in the aging process. A recent study (Nature Genetics, November2002) by the laboratory of Gary Ruvkun has shed light on the crucial changes in life span that occur when mitochondrial genes are removed from C.elegans. With the licensing of innovative mitochondrial transfection technology from the University of Virginia, Gencia Corporation is in a unique position to investigate the molecular interactions of mitochondrial DNA deletions, mitochondria and oxidative stress in human cells, and to bring the fruits of those investigations to the scientific community and the public. This Phase I SBIR proposal is directed toward the development and evaluation of human neural cell lines with several important features: the inducible production of specialized short hairpin loop RNAs that catalyze the degradation of the mitochondrial polymerase (PolG) RNA within cells, leading to a functional knockout of PolG, and a modified mitochondrial genome containing the 4977 bp "common deletion" that accumulates with aging. The specific aims outlined in this proposal detail the creation of these cell lines and the concomitant validation of the techniques used to create them, followed by a small-scale evaluation of their phenotypic responses to two important anti-oxidant compounds, vitamins E and C. A follow-on Phase II SBIR proposal will expand the library of compounds that are screened with the cell lines to include neurotrophic factors as well as novel antioxidant and neuroprotectiv compounds, developed either by Gencia Corporation or by our strategic corporate and/or academic partner organizations. A subsequent Phase III SBIR would permit the commercialization of neuroprotective /antioxidant compounds identified in Phase II for usage as therapies for neurodegenerative and other diseases. This project will represent a major step forward in the rational targeting and evaluation of compounds for therapeutic usage for a variety of human diseases.